An Analytical Study Of Pro: Khuda-Bukhush’s ‘Genetic Modulation Hypothesis’ Of Homeopathy

A.R. Khuda-Bukhsh, PhD., is a professor at the Cytogenetics and Molecular Biology Laboratory, Department of Zoology, University of Kalyani, Kalyani-741235, India. He has conducted groundbreaking investigations on homeopathic medicine since 1980 and has published dozens of scientific articles on homeopathic topics and other areas of research.

Khuda-Bukhsh  has published nearly thirty scientific articles in peer-reviewed  medical journals related to, or directly investigating homeopathic treatment, including cancer treatment, and yet many homeopathic practitioners  have  never  heard  of  him.  His  investigations  have  shown  strong  evidence  that  homeopathic  treatment  has  a biological effect, a fact that is still not widely accepted among scientists.

On the basis of his studies, Khuda-Bukush has also proposed a “hypothesis” regarding the biological mechanism of homeopathic cure. According to him, homeopathic drugs act by “modulating genetic expression”. Here, I am trying to evaluate his “hypothesis” on the basis of  an analytical study of his research works themselves.

Research works of Pro: Khuda Bukhsh play a great role in understanding the BIOLOGICAL EFFECTS of high dilution drugs, which is very important in scientific understanding of homeopathy.

But his works are not concerned about the fundamental questions of homeopathy such as ACTIVE PRINCIPLES of high dilution drugs, BIOLOGICAL MECHANISM of therapeutic actions of of high dilution drugs, MOLECULAR PROCESS involved in potentiation, and scientific explanation of SIMILIA SIMILIA CURENTUR. With out answering these fundamental questions, nobody can REDEFINE HOMEOPATHY scientifically and rationally. The greatness of the observations and contributions of Khuda Bukhsh is displayed only when they are interpreted in the light of MIT.

 

In the present article, I am trying to evaluate the ‘hypothesis’ proposed by Prof Khuda Bukush, through a critical appreciation of his EIGHT important scientific studies.

STUDIES CONDUCTED BY PROF. KHUDA BUKUSH, ON THE BASIS OF WHICH HE CLAIMS TO PROPOSE HIS ‘GENETIC MODULATION’ HYPOTHESIS REGARDING BIOLOGICAL MECHANISM OF HOMEOPATHIC CURE. MY COMMENTS ARE GIVEN UNDER EACH STUDY:

KHUDA-BUKHSH STUDY 1. Mechanism of biological action(s) of the potentized homeopathic remedies can be scientifically explained through a working hypothesis- By Professor A. R. Khuda-Bukhsh, PhD in Genetics, Former Head, Department of Zoology, University of Kalyani, India

One of the main reasons for the lack of acceptance of homeopathy as a scientific mode of treatment is the absence of a clear understanding of its mechanism of action, particularly of the ultra-high diluted potentized remedies. At the present state of our knowledge, the mechanism of action of the potentized homeopathic remedies diluted beyond Avogadro’s limit is extremely difficult to understand and explain. But here is a working hypothesis that can throw light on the possible mechanism of biological action of the potentized homeopathic remedies with concrete supportive evidences. In the beginning, let me focus on the intricacies that come in the way of a scientific explanation of the mechanism of action of the homeopathic remedies, particularly those diluted above the Avogadro’s limit. Firstly, to understand the mechanism of action of the potentized homeopathic drugs, one has to satisfactorily answer the problem of how the medicinal property of the original drug substance can be transferred to and retained by the ethanolic vehicle. Several working hypotheses have been proposed in this aspect (Anaganostatos et al., 1998; Allegre et al., 1989; Bellavite and Signorini, 2002; Elia and Niccoli 1999; Lo, 1996; Anick and Ives, 2007; Rey et al., 2007). The leading current proposal for the mode of action of such “ultra molecular” dilutions is that water (and aquatic ethanol as well) is capable of storing information relating to substances with which it has been in contact and subsequently can transmit this information to pre-sensitized biological system. The process is believed to be mediated by structural modification of water (or the aquatic ethanol, the “vehicle” of the homeopathic remedies), analogous to storage of information by magnetic media. Such information is retained in physical, rather than chemical form (Bellavite and Signorini, 2002). Several aspects of the “water structure” and “memory of water” have been critically analyzed recently by Chaplin (2007). Results of studies conducted on nature of molecular clustering in water solutions showed that as a solution is made more and more dilute, larger and very stable aggregates develop (Samal and Geckeler, 2001). This would mean that residual molecular clusters of the original substance may be present in homeopathic dilutions. Interestingly, alcohol also forms clusters with water (Wisnieski et al., 2001), which would further support the homeopathic analogy. Being an associative liquid, that is having hydrogen bonds between molecules, it is thus possible that alcohol can form its very own clusters. In fact, the presence of alcohol in water may actually be favorable for the moderation of succussion energy (by increasing vapor pressure). Andersson et al (1997) and Markovic et al (1999) were able to create individual clusters, using sudden evaporative cooling. According to these authors, cluster mixtures can generate many different isomeric forms in different geometric forms, and each isomer can represent a unique bit of information. Thus it is possible that millions of different “information bits” can be produced which may be “biologically active”. Hameroff and Penrose (2001) have proposed that microtubules in our brain are the seat of our conscious mind and consist of quantum micro switches (protein qubits), which contain “pure” ordered water. The microtubules interconnect every cell in a multicellular organism possibly being the means to extend the brain’s consciousness throughout the entire body. These authors further suggest that this microtubule network can explain the mind-body interactions, such as psycho-somatic pain. There are many research articles that suggest the role of cell cytoskeleton in genome regulation in cancer. Further there are evidences that support the fact that internal microtubules of the cell control mitosis (cell division), regulate the genes, decide which gene to turn on and so forth not only in terms of differentiation in development but also in health and in the steady-state. They believe that consciousness, the microtubules and quantum coherence play an essential role in health and diseases. Thus, in fine, the water cluster model and microtubule network model are comparable with Hahnemann’s concept of vital force in which a biological network offers all the action/reaction properties. Additionally, Hameroff’s (2001) work suggests that the quantum state of the microtubule’s elemental building block can be altered by an ‘ordered’ water structure attached to the external side of the microtubules. The water cluster(s), which is (are) specific to the problems, can simply throw the right switch or switches to correct the error in the network. Thus molecular architecture of water has a key role to play in understanding homeopathic mechanism of action and has therefore been vigorously studied (Dantas et al., 2007; Elia et al., 2007; Anick and Ives (2007).

The next part of the mechanism that should be addressed is how the tiny drop of a potentized remedy or a few sugar globules soaked with the remedy can trigger the recovery process of the disease/abnormal symptoms. There are several hypotheses based on in vivo and in vitro studies with various animal models in relation to various immunological, cardiovascular and molecular aspects and various kinds of hypotheses like “hormesis”, “allergic reaction”, or “immunological responses” etc. have been proposed (see The HomBRex database, available at: http://www.carstens-stiftung.de/hombrex ). These hypotheses have their apparent merits, but also suffer from some inadequacies in explaining the action of homeopathic drugs in unicellular organisms like protozoans or bacteria. Any acceptable hypothesis has to be able to explain the mechanism of action universally in all kinds of organisms where homeopathic remedies have been reported to act. Khuda-Bukhsh (1997, 2003, 2006, 2009) proposed a hypothesis based on various scientific evidences that potentized homeopathic drugs act through regulation of relevant gene expression. The possible pathways and sites of action have also been discussed by this group (Khuda-Bukhsh, 1997; Mallick et al., 2003; Pathak et al., 2006; 2007; Khuda-Bukhsh and Pathak, 2008). According to this hypothesis, homeopathic remedies carry specific “signals”/”information” that can be identified by specific receptors of the cells. These “signals” can act as a “trigger” for turning “on” or “off” some relevant genes, initiating a cascade of gene actions to alter and correct the gene expressions that went wrong to produce the disorder/disease. Presumably, the homeopathic drugs are mediated through cytokine responses. Thus, administration of a potentized homeopathic drug can elicit response in suitable signal proteins and can either up-regulate or down-regulate such signal proteins to bring back the recovery of the patient to normal health (Khuda-Bukhsh et al., 2009). For the same reason a global microarray after a homeopathic drug administration in a disease/abnormal state should provide evidence for modulated expression of a large number of genes that has actually been reported recently (D Oliveira et al., 2008).

In higher eukaryotes like mammals, the regulation of gene expression is, however, a very complex phenomenon. In principle, the expression of genes might be regulated at any one of several stages. At least five control points can be distinguished that form the series, through which gene expression is regulated (see Lewin 2004): Activation of structure – Initiation of transcription – Processing the transcript – Transport to cytoplasm – Translation of mRNA. Many eukaryotic genes have multiple regulatory binding sites and are controlled by more number of regulatory proteins (see Watson et al., 2004). Enhancers bind regulators responsible for activating the gene at a given time and place. Alternative enhancers bind different groups of regulators and control expression of the same gene at different times and places in response to different signals. Some regulators bind sites far from the genes they control, by repressor proteins. Repressors work by a variety of ways, including the way which is called “gene silencing”. Modification to regions of chromatin keep genes in sometimes large stretches of DNA “switched off”. Similarly, there are “activators”, which do not always have well-defined structure. Activators recruit the transcriptional machinery to the gene, and sometimes recruit nucleosome modifiers that help the transcription machinery bind at the promoter. Activators are known to work synergistically to integrate signals. Synergy is critical for signal integration by activators. Each signal is communicated to the gene by a separate activator (signal recognition particle). Signals are often communicated to transcriptional regulators through signal transduction pathways. There are various ways that signals are detected by a cell and communicated to a gene. It can also refer to the “information” as it passes from detection of the ligand to the regulators that directly control the genes- that is, as it passes along a signal transduction pathway. In a signal transduction pathway, the initiating ligand is typically detected by a specific cell surface receptor: the ligand binds to an extracellular domain of the receptor and this binding is communicated to the intracellular domain. From there the signal is relayed to the relevant regulator, often through a cascade of kinases. Generally binding of ligand alters the shape (and thus activity) of the intracellular domain. Alternatively the ligand can act simply to bring together two or more receptor chains, allowing interaction domains of those receptors to activate each other (see Watson et al., 2004). However, how a homeopathic drug can elicit response in a cell receptor and can bind with the receptor is not precisely known. These are the areas that should be more specifically searched. Our research points out a possibility that homeopathic drug can have interaction with some nano-particles that can tag onto some proteins. That can make it possible for some drug-associated nano-particles to get attached to some proteins, which then act as the ligand (Bhattacharyya et al., 2008). However, this is only speculative at this stage of our knowledge and needs validation by further research. A recent finding that nano-particles of metals can be traced in ultra-high diluted homeopathic remedies through high powered electron microscopes vindicates our hypothesis.

Likewise, there are STAT and MAP kinase pathways activated by ligand (a cytokine) in which phosphorylation and dephosphorylation of different kinases occur to carry forward the information to produce activities in genes downstream as a cascade of reactions. Perhaps signals of potentized homeopathic drugs are carried through this method (Khuda-Bukhsh, 2009; Khuda-Bukhsh et al., 2009). Gene expression can also be controlled by “signals” received by a cell from its environment. For example, the presence of sugar lactose activates the transcription of the lac operon in E. coli, while viral infection activates the expression of β-interferon gene in mammals (see Watson et al., 2004). Interestingly, in our recent experiment on E. colisubjected to low dose of arsenic, Arsenicum Album 30C showed modulatory effect in their glucose uptake (results unpublished). Generally speaking, the strategies that are used to instruct genetically-identical cells to express distinct sets of genes and thereby perform different sets of function include m-RNA localization, cell-to-cell contact and signaling through the diffusion of a secreted signaling molecule. Microarray assays can thus help further in the analysis of gene expression profiles of experimental organisms administered micro doses of the homeopathic drug.

In fine, the reasons and circumstantial evidences that support gene regulatory hypothesis are:

  • The use of various state-of-the art techniques has now quite convincingly proved the efficacy of several potentized homeopathic drugs diluted beyond Avogadro’s limit in modulating significantly many parameters of study. These parameters are considered scientifically acceptable and dependable. The potentized homeopathic remedies are capable of modulating such parameters, while the succussed alcohol (placebo) can not. Further, the homeopathic drug can bring about changes in multiple parameters at any given time frame, which without a cascade of gene actions is not possible.
  • The homeopathic remedies are diluted to such an extent that there can not be physical existence of a single molecule of the original drug substance. Therefore it is simply not possible for the remedy to react chemically with all the cells undergoing some degree of quantifiable/visible changes at the patho-physiological level. But there are many evidences that demonstrate modulations by the potentized homeopathic remedy (diluted beyond Avogadro’s limit) in regard to almost every endpoint studied.
  • This leaves us with the only possibility that the homeopathic drug carries molecular imprints or definite signal/information of the original drug molecule that can be deciphered by the cells’ receptors. If one knows a particular language, no matter if the size of the script is small or large, the message will carry the same meaning. Therefore, if they indeed carry some signals, and the cells can perceive that, then there should be a reactive process going on in the transduction of the signal to the target organ. Most of the parameters of study, particularly the expression of signal proteins, are indicators of the fact that their genes have been expressed in terms of either up-regulation or down regulation, as the case may be, that is good evidence of their ability to exert influence on the regulatory genes of these signal proteins; since the expression of these signal proteins and some other specific receptor proteins (like AhR or PCNA) have also been tested positive by some other confirmatory tests like immunohistochemical localization as well. Further, the ability of the homeopathic drugs in bringing about structural alterations at the sub-cellular levels in some vital organs (like liver) has been evidenced by electron microscopic studies as well, that would also speak for its ability to tackle matters at the molecular level.
  • The use of several modern techniques involving monoclonal antibodies for the detection of these signal proteins through immunoblot experiments are currently accepted to be quite dependable scientific tool for ascertaining gene modulating effects of any drug.
  • Nanoparticles have been shown to have effect on the physico-chemical property of the homeopathic dilutions (Bhattacharyya et al., 2008), that may indicate some role of the nano-particles during the potentization process of the homeopathic drugs.
  • All parameters of study tested so far are strictly gene controlled.
  • Homeopathic dilutions have been noted to have demonstrable and reproducible action on plants, and bacteria, which lack any central nervous system.
  • And finally, the simultaneous treatment of Actinomycin D, a transcription blocker, the action of a potentized homeopathic remedy fails (Dutta et al., 1999; Chakrabarti et al., 2001).

Thus, our extensive research on homeopathy yielded much suggestive information for understanding the biological part of the mechanism of action. More in-depth research, particularly by other independent researchers, to test and verify our research findings will be highly solicited, because validation of results obtained by us can indeed open up newer avenues, paving the way for a better understanding of the mechanism and pathways of biological action of the potentized homeopathic remedies in near future, and to establish the “gene regulation” hypothesis as universally acceptable one.

 My comments on Study 1:

I really wonder what scientist of big stature like him really intended by including in his articles some introductory references to the pseudo scientific theories and concepts of Anaganostatos et al., 1998; Allegre et al (1989) Bellavite and Signorini (2002), Elia and Niccoli  (1999),  Lo (1996), Anick and Ives (2007) , Rey et al (2007), Beneviste  and Signorini (2002),  Chaplin (2007), Samal and Geckeler (2001), Wisnieski et al (2001),  Andersson et al (1997), Markovic et al (1999)  Hameroff and Penrose (2001) etc etc. All these pseudo-scientific references severely damage the scientific credentials of the hypothesis proposed by the professor.

Professor Khuda Bukhush rightly says:

“One of the main reasons for the lack of acceptance of homeopathy as a scientific mode of treatment is the absence of a clear understanding of its mechanism of action, particularly of the ultra-high diluted potentized remedies. At the present state of our knowledge, the mechanism of action of the potentized homeopathic remedies diluted beyond Avogadro’s limit is extremely difficult to understand and explain”.

After saying this much, he offers us to propose “a working hypothesis that can throw light on the possible mechanism of biological action of the potentized homeopathic remedies with concrete supportive evidences”.

He identifies the primary task of such a hypothesis: “Firstly, to understand the mechanism of action of the potentized homeopathic drugs, one has to satisfactorily answer the problem of how the medicinal property of the original drug substance can be transferred to and retained by the ethanolic vehicle.”

The sad part of his neatly written article is that no where he tries “answer the problem of how the medicinal property of the original drug substance can be transferred to and retained by the ethanolic vehicle”.  Instead, he quotes from various references from pseudoscientific literature, never explaining his stand on those ideas he quoted.

Listen to some of his quotes:

“The leading current proposal for the mode of action of such “ultra molecular” dilutions is that water (and aquatic ethanol as well) is capable of storing information relating to substances with which it has been in contact and subsequently can transmit this information to pre-sensitized biological system.” Does the professor authenticate this “leading current proposal”? He keeps silent.

The process is believed to be mediated by structural modification of water (or the aquatic ethanol, the “vehicle” of the homeopathic remedies), analogous to storage of information by magnetic media”. Author says “process is believed to be”. Does he share this “belief”?

“Such information is retained in physical, rather than chemical form (Bellavite and Signorini, 2002).” What is your opinion on this, sir?

Several aspects of the “water structure” and “memory of water” have been critically analyzed recently by Chaplin (2007).” Do you agree with Chaplin’s analysis?

“Results of studies conducted on nature of molecular clustering in water solutions showed that as a solution is made more and more dilute, larger and very stable aggregates develop (Samal and Geckeler, 2001). This would mean that residual molecular clusters of the original substance may be present in homeopathic dilutions.” Here also, professor says “may be”. He has no his own opinion.

“Interestingly, alcohol also forms clusters with water (Wisnieski et al., 2001), which would further support the homeopathic analogy. Being an associative liquid, that is having hydrogen bonds between molecules, it is thus possible that alcohol can form its very own clusters. In fact, the presence of alcohol in water may actually be favorable for the moderation of succussion energy (by increasing vapor pressure).” See-  again he says “it is possible” and “may actually be favorable”!

“Andersson et al (1997) and Markovic et al (1999) were able to create individual clusters, using sudden evaporative cooling. According to these authors, cluster mixtures can generate many different isomeric forms in different geometric forms, and each isomer can represent a unique bit of information. Thus it is possible that millions of different “information bits” can be produced which may be “biologically active”. Professor says “according to these authors”. He does not open up his mind here also.

Hameroff and Penrose (2001) have proposed that microtubules in our brain are the seat of our conscious mind and consist of quantum micro switches (protein qubits), which contain “pure” ordered water. The microtubules interconnect every cell in a multicellular organism possibly being the means to extend the brain’s consciousness throughout the entire body. These authors further suggest that this microtubule network can explain the mind-body interactions, such as psycho-somatic pain. There are many research articles that suggest the role of cell cytoskeleton in genome regulation in cancer. Further there are evidences that support the fact that internal microtubules of the cell control mitosis (cell division), regulate the genes, decide which gene to turn on and so forth not only in terms of differentiation in development but also in health and in the steady-state. They believe that consciousness, the microtubules and quantum coherence play an essential role in health and diseases”. Listen, professor has no opinion- he said “they believe”!

“Thus, in fine, the water cluster model and microtubule network model are comparable with Hahnemann’s concept of vital force in which a biological network offers all the action/reaction properties”.  Here the professor seems to agree with the idea that “water cluster model and microtubule network model are comparable with Hahnemann’s concept of vital force”!

“Additionally, Hameroff’s (2001) work suggests that the quantum state of the microtubule’s elemental building block can be altered by an ‘ordered’ water structure attached to the external side of the microtubules. The water cluster(s), which is (are) specific to the problems, can simply throw the right switch or switches to correct the error in the network.” OK sir, “Hameroff’s work suggests”- but what is your opinion on that “suggestion”?

He concludes this inquiry by the statement: “Thus molecular architecture of water has a key role to play in understanding homeopathic mechanism of action and has therefore been vigorously studied (Dantas et al., 2007; Elia et al., 2007; Anick and Ives (2007).”

Did this discussion by the learned professor positively contribute any thing for the scientific understanding of the “mechanism of action of the potentized homeopathic drugs, one has to satisfactorily answer the problem of how the medicinal property of the original drug substance can be transferred to and retained by the ethanolic vehicle”, as he initially identified as the primary task of a hypothesis regarding biological mechanism of homeopathic drug actions?

Leaving the “first part” in utter confusion, the professor moves on to “second part” of his ‘hypothesis’:

“The next part of the mechanism that should be addressed is how the tiny drop of a potentized remedy or a few sugar globules soaked with the remedy can trigger the recovery process of the disease/abnormal symptoms”

In this article, Khuda-Bukhsh explains his ‘hypothesis’ that potentized homeopathic drugs act through “regulation of relevant gene expression”, which is claimed to be based on various “scientific evidences” he could collect through his scientific studies.

He says: “According to this hypothesis, homeopathic remedies carry specific “signals”/”information” that can be identified by specific receptors of the cells. These “signals” can act as a “trigger” for turning “on” or “off” some relevant genes, initiating a cascade of gene actions to alter and correct the gene expressions that went wrong to produce the disorder/disease. Presumably, the homeopathic drugs are mediated through cytokine responses. Thus, administration of a potentized homeopathic drug can elicit response in suitable signal proteins and can either up-regulate or down-regulate such signal proteins to bring back the recovery of the patient to normal health”.

What does this statement show? It shows he has no any idea about the exact active principles of potentized drugs, which should actually be the basis of any rational ‘hypothesis’ regarding the ‘biological mechanism’ of drug actions. He says “homeopathic remedies carry specific “signals”/”information”. What is the mechanism by which these “signals/information” are transferred from drugs substances into potentizing medium during the process of potentization? In what form these  “signals” exist in potentized drugs? In what form these “information” is stored, and transmitted? Is he talking about “electromagnetic signals” or ‘waves’ as our ‘energy medicine’ theoreticians talk about? Or, is it “chemical” signals or ‘nanoparticles’ as he proposes in later part of his article? How can an eminent biologist can talk about active principles of drug substances and their biological actions in such vague terms of  “signals/information”? How can he say these “signals/information” being “identified by specific receptors of the cells”? Without giving any clarification regarding the form in which these “signals/information” exist, how can he say these ‘signals” can act as a trigger for turning “on” or “off” some relevant genes, initiating a cascade of gene actions to “alter and correct the gene expressions that went wrong to produce the disorder/disease”?

What is the role of “similia similibus curentur” in this process? How can anybody make a hypothesis about homeopathy, without any mention of its fundamental principle of ‘similia similibus curentur’? How can he incorporate  this concept of “up-regulate or down-regulate such signal proteins to bring back the recovery of the patient to normal health” into the frame work of ‘similia similibus curentur’?

According to the author, “administration of a potentized homeopathic drug can elicit response in suitable signal proteins and can either up-regulate or down-regulate such signal proteins to bring back the recovery of the patient to normal health”. Where as according to him, “potentized homeopathic drugs” mean “signals/information’, hat is the proposed biochemical process by which the “signals” “elicit response in suitable signal proteins”? He is expected to explain. According to homeopathy, only indicated drugs can produce any therapeutic effect. As such, Professor should explain how the biological actions of “signals” of indicated drugs differ from non-indicated drugs? All the studies of the author no where consider or mention about the importance of indicated remedy, which very important in homeopathy.

He continues: “However, how a homeopathic drug can elicit response in a cell receptor and can bind with the receptor is not precisely known. These are the areas that should be more specifically searched”. That means, his ‘hypothesis’ does not address the fundamental aspect of ‘biological mechanism’ of homeopathic cure!

Listen to his another statement: “Our research points out a possibility that homeopathic drug can have interaction with some nano-particles that can tag onto some proteins. That can make it possible for some drug-associated nano-particles to get attached to some proteins, which then act as the ligand (Bhattacharyya et al., 2008). However, this is only speculative at this stage of our knowledge and needs validation by further research. A recent finding that nano-particles of metals can be traced in ultra-high diluted homeopathic remedies through high powered electron microscopes vindicates our hypothesis.” He is now talking about “possibilities”! See him conveniently shifting to “nanoparticle” theory from his “signal/information” theory he originally proposed! How can he say his hypothesis is “vindicated” by the “recent finding that nano-particles of metals can be traced in ultra-high diluted homeopathic remedies through high powered electron microscopes”? What will happen to this “vindication” if “nanoparticle” theory of IIT-B scientists are proved wrong?

He next jumps to another theory: “Likewise, there are STAT and MAP kinase pathways activated by ligand (a cytokine) in which phosphorylation and dephosphorylation of different kinases occur to carry forward the information to produce activities in genes downstream as a cascade of reactions. Perhaps signals of potentized homeopathic drugs are carried through this method”. It is “perhaps”- he is not sure about anything he says. His ‘hypothesis’ is all about various ‘possibilities’, ‘probable’, ‘may have’  and ‘perhaps’. Is it the way a scientist should propose a ‘scientific hypothesis’ to explain a controversial phenomenon?

KHUDA-BUKHSH STUDY 2. Evidence in support of gene regulatory hypothesis: Gene expression profiling manifests homeopathy effect as more than placebo . By Santu Kumar Saha, Sourav Roy and Anisur Rahman Khuda-Bukhsh

  ABSTRACT

 Background: Use of ultra-high diluted remedies in homeopathy and their claimed efficacy in curing diseases has been challenged time and again by non-believers despite many evidence-based positive results published in favor of their efficacy in curing/ameliorating disease symptoms.

Aims: To test the ability of ultra-high diluted homeopathic remedies beyond Avogadro’s limit, if any, in manifesting gene modulating effects in controlled in vitro experimental model.

Methods: Since cancer cells manifest aberrant epigenetic gene expressions, we conducted global microarray gene expression profiling of HeLa cells (an established epigenetic model of HPV18 positive cell line) treated with two different potentized homeopathic remedies, namely, Condurango 30c and Hydrastis canadensis 30C (used in the treatment of cancer), as compared to that of placebo (succussed alcohol 30c).

Results: Data revealed distinctly different expression patterns of over 100 genes as a consequence of treatment with both homeopathc remedies compared to placebo.

Conclusion: Results indicate that action of the potentized drugs was “more than placebo” and these ultra-highly diluted drugs acted primarily through modulation of gene expression.

 Key words: Gene regulatory hypothesis, DNA microarray profile, potentized remedies.

 Introduction: Homeopathy is one of the most widely practiced controversial modes of treatment as it uses extremely diluted remedies in micro-doses to alleviate patients’ complaints. Despite being used for centuries with great effect, non-believers of homeopathy mainly ask two questions: (a) as drugs diluted beyond Avogadro’s limit (6.023 x 1023) are not expected to contain even one single molecule of the original drug substance, how can the homeopathic medicines then be physical-chemically different from “vehicle” ethanol, and claimed to be effective in curing many diseases? (b) What might the mechanism of action of these ultra-highly diluted remedies be?

In this report, we present gene expression profiling results that give strong support to a hypothesis proposed by Khuda-Bukhsh [1, 2], to wit, that potentized homeopathic drugs act through regulation of gene expression. Thangapazham et al. [3], in turn, apparently failed to find changes in gene expression induced by homeopathic drugs used in in vitro experiments with human breast cancer cells MDA-MB-231, human prostate cancer cells DU-145 and LNCaP, and rat prostate cancer cells MAT-LyLu treated with some potentized homeopathic remedies like Conium maculatumSabal serrulataThuja occidentalisAsterias rubens, and Phytolacca decandra in dilutions 30c, 200c and 1,000c, and Carcinosinum (1,000 c) although they Int J High Dilution Res 2013; 12(45):162-167 163  had found in an earlier in vivo study reduction in the number and size of cancer nodules supported by histological analysis in prostate cancer MAT-LyLu cell–injected Copenhagen rats given homeopathic treatment containing ThujConSabal serrulata, and in MAT-LyLu cell with Carc [4]. However, the authors did not conduct gene expression studies of any signal proteins in these rats in vivo, where modulation of certain gene expression would be expected.

The controversy surrounding research in homeopathy moves along two different directions. On the one hand, the mechanism of action of the potentized homeopathic remedies diluted beyond Avogadro’s limit remains an open research problem, which has received widespread attention from the research community, and many competing hypotheses have been proposed. On the other hand, some studies that indicate homeopathy is no better than placebo, which thus makes any claim about the mechanism of action of homeopathic drugs null and void. There have been many results highly critical of homeopathy. For example, The Lancet of August 27, 2005 featured an article that was highly dismissive of homeopathy [5] along with a press release: “homeopathy is no better than placebo”. The meta-analysis was accompanied by a short, anonymous editorial entitled ‘The end of homoeopathy’. This resulted in the eruption of a ravaging controversy: whether homeopathy is really better than placebo or not, culminating in a debate in the British Parliament [6]. Homeopathy was decried and its use as a beneficial and scientifically proven mode of treatment was voted against. However, most clinical research conducted with homeopathic medicines has shown positive results [7]. Biological activities of ultra-high diluted potentized homeopathic remedies have also been confirmed by basic research studies with respect to a multitude of scientifically accepted protocols, both in animals and plants in vivo and in vitro [8]. Nevertheless, the argument persisted, as ultra-highly diluted homeopathic medicines are supposed to possess “nothing” in terms of the original drug molecules, and as such their effect on biological systems was unacceptable until recently. During the last few years, presence of “nanoparticles” of the starting materials, even at extremely high dilutions, has been reported [9, 10]. Moreover, Montagnier [11] countered through experiments that “high dilutions of something are not ‘nothing’, they are water structures which mimic the original molecules”.

We decided to use one of the most modern tools, global microarray profiling of genes, to verify whether the effects of ultra-highly diluted homeopathic drugs on the expression profile of genes were distinctly different compared to “placebo”. DNA microarrays are widely used to measure the expression levels of large numbers of genes simultaneously, with the aid of selective probes under highly stringent conditions. Gene expression profiling experiments are specially effective for monitoring the expression levels of thousands of genes to study the effects of certain treatments, for example, to identify genes whose expression changes in response to pathogens or drugs, by comparing the gene expression in the infected and drug-treated cells or tissues [12].

Materials and methods: The aim of the present study was, therefore, to find out whether the global gene expression profiles in cancer cell line, HeLa (an established epigenetic model of HPV18 positive cell line) differed significantly, in quantitative as well as qualitative terms, after treatment with 2 homeopathic remedies used against cancer [13-15], namely, Condurango 30c and Hydrastis canadensis 30c (both diluted 10-60 times, much above Avogadro’s limit) compared to ethanol 30c, (placebo, prepared from the same stock of 70% alcohol giving 10 jerks at each step in the same manner as the verum was potentized). Affymetrix Gene Chip Human Primeview gene expression arrays were used for this purpose.

Cervical cancer cell line HeLa was obtained at National Centre for Cell Science, Pune. The cells were routinely maintained in Dulbecco’s Modified Eagle’s Medium supplemented with 10% FBS and 1% antibiotic at 37 ºC in a humidified incubator containing 5% CO2. The cells were plated one day before treatment. 4% (v/v) of drugs and “placebo” (succussed ethanol 30c, 10 jerks at each step of dilution) were added to the cell Int J High Dilution Res 2013; 12(45):162-167 164 cultures and kept for 48 hr. Cells without any treatment in the normal medium were considered as negative control.

Separate groups of cells were subjected to treatment with Cond 30c, Hydr 30c, and succussed alcohol placebo. The cells were sent to iLife Discoveries, Gurgaon, India for providing us global microarray data conducted on Affymetrix platform, using 25-mer probes. The total number of probes detected for the experiment was 49,395; hybridization was performed at 45 oC for 16 hrs at 60 rpm.

Slides were scanned with 3000 7G microarray scanner and raw data sets were extracted from the Cel (raw intensity) files. Microarray data analysis, differential gene expression analysis, fold change analysis and cluster analysis were performed using GeneSpring GX12.5 software.

 Results and discussion:  Microarray data analyses showed that out of a total of 40,678 genes, for which probe sets remained after data processing, normalization and quality control, 6,024 were differentially expressed at a p-value cutoff of 0.05, in a one-way ANOVA. The expression levels of the genes in the cells were treated with Hydr 30c (Set I) and Cond30c (Set II) were compared to the levels in the untreated (control) as well as succussed ethanol 30c (placebo) treated cells (Set III). Using a cutoff of 1.5 folds it was observed that in Set I, 3 genes each were up- and down-regulated when compared to the control as well as Set III samples. Similarly, there were 2 genes in Set II that were up-regulated by at least 1.5 folds when compared to the two sets mentioned above, whereas 122 genes were down-regulated by ≥1.5 folds. Two and 10 genes were up- and down-regulated by ≥ 1.5 folds, respectively, in both sets (Set I and Set II) when compared to the control as well as Set III. In addition, there were another 23 genes in Set I and 12 genes in Set II that were differentially expressed by ≥ 1.5 folds, when compared to Set III. Upon comparing the expression levels of genes in Set I and Set II, it was observed that for 36 genes, there were ≥ 1.5 fold differences in expression between the 2 treatments. These findings suggest that the drugs did not only affect the gene expression, but also that the effect of one drug was different from the other in a number of genes.

The findings of the present study clearly demonstrate that the expression profiles of certain genes of the drug-treated HeLa cells were significantly different from that of the placebo treated cells. This suggests that both drugs and placebo differed in their ability to trigger gene responses, some of which were implicated in cancer. Thus, although the drugs were ultra highly diluted, they still retained the ability to trigger gene responses in a cascade of reactions, diving support to Khuda-Bukhsh’s hypothesis[1-2]. Epigenetic modifications are a hallmark of cancer, and a large number of genes remain in modified state of expression in cancer cells. Both Hydr[13, 15] and Cond [14, 15] had been previously reported to induce apoptosis in cancer cells, while “placebos” do not exhibit this property. Therefore, it is logical to infer that homeopathic remedies contain some form of molecular imprints of the original drug substance [16], while the “placebo”, in the absence of such imprints, fails to elicit the required effect.

Incidentally, ultra-highly diluted preparations of glucose 30c, Arsenicum album 30c, and Arnica montana 30c were shown to induce gene modulatory effects in bacteria, E. coli and yeast Saccraromyces cerevisiae, either subjected to insult with sodium arsenite or UV-irradiation [17-19]. Those authors [17-19] explained that the potentized homeopathic remedies carry specific “signals”/”information” that can be identified by certain cell receptors. Those “signals” may act as a “trigger” for turning “on” or “off” some relevant genes, initiating a cascade of gene actions, while the “placebo” failed to elicit any such favorable responses. It has been previously documented that administration of a potentized homeopathic drug altered the expressions of a large number of signal proteins in mice induced to develop skin cancer [20]. Int J High Dilution Res 2013; 12(45):162-167 165

The results of the present study, therefore, contribute to support the gene regulatory hypothesis proposed by Khuda-Bukhsh [1, 2]. Further studies are needed to ascertain the ability of Cond 30c, if any, to modulate activities like DNA methylation/demethylation and histone acetylation/deacetylation, 2 hallmarks of epigenetic modifications, to produce additional support to the gene regulatory hypothesis.

 My comments on  Study 2:

This is a great study, but the authors arrived at wrong conclusions. They actually wanted to “prove” their hypothesis regarding the biological mechanism of potentized homeopathic drug action in terms of “genetic modulation”. That is why they made wrong interpretations of this great work.

Actually, this study leads us to a different conclusion if analyzed rationally. This study has proved that the rightly selected potentized drugs can rectify the errors happened in normal gene regulation processes produced by the action of pathogenic molecules. Errors in genetic expression and dna replication are produced by the inhibition of various enzymes involved in essential molecular processes such as “DNA methylation/demethylation and histone acetylation/deacetylation”. Endogenous or exogenous pathogenic molecules bind to the enzymes and inhibit them, resulting in faulty genetic expression and dna replication. This is exactly how various carcinogenic substances produce tumor growth and cancers. Molecular imprints of these carcinogenic agents can act as artificial binding sites for these carcinogenic molecules, and remove the molecular inhibitions that produced errors in genetic expression and dna replication.

It should be specifically noted that these experiments were conducted by determining the effects of of potentized drugs in reducing or rectifying the in the “genetic expression aberrations” already produced by pathogenic agents. This study no way prove potentized drugs has any effect up on genetic expressions  in the absence of existing pathological “aberrations”. If the hypothesis regarding “genetic modulation” power of potentized drugs were right, it should be possible to produce “aberrations” by using potentized drugs upon healthy cells. This study proves that potentized drugs can act only if pathogenic molecules are present in the system, which in turn proves that potentized drugs act ONLY upon pathogenic molecules, and not upon normal biological molecules. Potentized drugs act not by genetic modulation, but by rectifying the errors in in genetic modulation produced by pathogenic molecules inhibiting the enzyme system.

In fact, all pathological conditions are not produced by modulation of genetic expression. There are many diseases that are caused by inhibitions of various enzymes, cellular receptors, transport molecules and various other biological molecules. Many pathogenic agents such as infectious agents, drugs, toxins and metabolic byproducts produce diseases by inhibiting different biological molecules other than those involved in genetic modulation.

I request respected KHUDA BUKHUSH to modify his ‘hypothesis’ a little: Potentized homeopathic drugs act by rectifying the molecular errors produced by pathogenic molecules. Potentized drugs can also bind to pathogenic molecules and rectify the errors  they produced in the enzymes involved in genetic expression and dna replication. All his reported great research studies will ratify such a rational hypothesis.

It is obvious that Prof Khuda- Bukush is in utter confusions regarding the active principles of potentized drugs, as well as the molecular mechanism of their biological actions. He says:

“Despite being used for centuries with great effect, non-believers of homeopathy mainly ask two questions: (a) as drugs diluted beyond Avogadro’s limit (6.023 x 1023) are not expected to contain even one single molecule of the original drug substance, how can the homeopathic medicines then be physical-chemically different from “vehicle” ethanol, and claimed to be effective in curing many diseases? (b) What might the mechanism of action of these ultra-highly diluted remedies be?”.

Read him further: “The controversy surrounding research in homeopathy moves along two different directions. On the one hand, the mechanism of action of the potentized homeopathic remedies diluted beyond Avogadro’s limit remains an open research problem, which has received widespread attention from the research community, and many competing hypotheses have been proposed. On the other hand, some studies that indicate homeopathy is no better than placebo, which thus makes any claim about the mechanism of action of homeopathic drugs null and void. There have been many results highly critical of homeopathy. For example, The Lancet of August 27, 2005 featured an article that was highly dismissive of homeopathy [5] along with a press release: “homeopathy is no better than placebo”. The meta-analysis was accompanied by a short, anonymous editorial entitled ‘The end of homoeopathy’. This resulted in the eruption of a ravaging controversy: whether homeopathy is really better than placebo or not, culminating in a debate in the British Parliament [6]. Homeopathy was decried and its use as a beneficial and scientifically proven mode of treatment was voted against. However, most clinical research conducted with homeopathic medicines has shown positive results [7]. Biological activities of ultra-high diluted potentized homeopathic remedies have also been confirmed by basic research studies with respect to a multitude of scientifically accepted protocols, both in animals and plants in vivo and in vitro [8]. Nevertheless, the argument persisted, as ultra-highly diluted homeopathic medicines are supposed to possess “nothing” in terms of the original drug molecules, and as such their effect on biological systems was unacceptable until recently. During the last few years, presence of “nanoparticles” of the starting materials, even at extremely high dilutions, has been reported [9, 10]. Moreover, Montagnier [11] countered through experiments that “high dilutions of something are not ‘nothing’, they are water structures which mimic the original molecules”.

Kindly notice his confusions in this regard: “During the last few years, presence of “nanoparticles” of the starting materials, even at extremely high dilutions, has been reported. Moreover, Montagnier  countered through experiments that “high dilutions of something are not ‘nothing’, they are water structures which mimic the original molecules”.  That means, he has no any idea of molecular imprinting involved in homeopathic potentization. He relies up on “nanoparticle” theory as well as Montainer’s observations of  “water structures which mimic the originalmolecules”.

Had Prof. Khuda Bukush arrived at molecular imprints as active principles of potentized drugs, he could have utilized his wonderful research works for providing a rational and scientific explanation for the biological mechanism of homeopathic therapeutics, which he actually could not. I feel extremely sorry for that.

KHUDA-BUKHSH STUDY 3. Comparative efficacy of two microdoses of a potentized homoeopathic drug, Cadmium Sulphoricum, in reducing genotoxic effects produced by cadmium chloride in mice: a time course study- By Datta SSMallick PPRahman Khuda-Bukhsh AA.

“In this experiment, the researchers tested  the efficacy of two potencies of a homeopathic drug, Cadmium Sulphoricum (Cad Sulph), in reducing the genotoxic effects of Cadmium chloride in mice.  For this, healthy mice, Mus musculus, were intraperitoneally injected with 0.008% solution of CdCl2 @ 1 ml/100 gm of body wt (i.e. 0.8 mcg/gm of bw), and assessed for the genotoxic effects through such studies as chromosome aberrations (CA), micronucleated erythrocytes (MNE), mitotic index (MI) and sperm head anomaly (SHA), keeping suitable succussed alcohol fed (positive) and CdCl2 untreated normal (negative) controls. The CdCl2 treated mice were divided into 3 subgroups, which were orally administered with the drug prior to, after and both prior to and after injection of CdCl2 at specific fixation intervals and their genotoxic effects were analyzed. While the CA, MNE and SHA were reduced in the drug fed series as compared to their respective controls, the MI showed an apparent increase. The combined pre- and post-feeding of Cad Sulph showed maximum reduction of the genotoxic effects. Both Cad Sulph-30 and 200 were able to combat cadmium induced genotoxic effects in mice and that combined pre- and post-feeding mode of administration was found to be most effective in reducing the genotoxic effect of CdCl2 followed by the post-feeding mode.”

 My comment on Study 3:

What did this experiment actually proved? It proved that cadmium sulph in crude form can act as a carcinogen as shown by increased  chromosome aberrations (CA), micronucleated erythrocytes (MNE) and sperm head anomaly (SHA). Potentized cadmium sulph could prevent, reduce or rectify these damages caused by crude cadmium sulph. We do not know the exact molecular mechanism by which cadmium sulph molecules produce genotoxic effects. Probably, cadmium sulph might have produced molecular inhibitions upon various enzymes involved in the process of genetic expression. Molecular imprints of cadmium sulph contained in its potentized form could bind to the cadmium sulph molecules and relieve the enzyme system from their inhibitions. This experiment no way prove potentized drugs “ACT” by  “interacting with DNA”- it only proved potentized drugs can prevent dna damages caused by the inhibitions of genetic expression enzyme systems produced by pathogenic molecules.

KHUDA-BUKHSH STUDY 4. Efficacy of the potentized homeopathic drug, Carcinosin 200, fed alone and in combination with another drug, Chelidonium 200, in amelioration of p-dimethylaminoazobenzene-induced hepatocarcinogenesis in mice- By Biswas SJPathak SBhattacharjee NDas JKKhuda-Bukhsh AR.

“This study was conducted to examine whether the potentized homeopathic remedy Carcinosin 200, fed alone and in combination with Chelidonium 200, has differential protective effects against p-dimethylaminoazobenzene (p-DAB)-induced hepatocarcinogenesis in mice. Liver tumors were induced in mice through chronic feeding of p-DAB (initiator) and phenobarbital (PB, promoter). The mice were divided into two subgroups: (1) one was fed potentized Alcohol 200 and served as controls; and (2) the other was fed Carcinosin 200 alone or in combination with Chelidonium 200 and divided into several sets. The relative efficacy of the two potentized remedies, alone or in combination, in combating hepatocarcinogenesis was assessed through several cytogenetical endpoints such as chromosome aberrations, induction of micronuclei, sperm head anomaly, and mitotic index at several intervals of fixation  Several toxicity biomarkers such as acid and alkaline phosphatases, glutamate oxaloacetate transaminase, glutamate pyruvate transaminase, and lipid peroxidation activity were also assayed in three organs of treated and control mice. In addition, recovery by the homeopathic drugs, if any, of tissue damage inflicted because of chronic feeding of p-DAB and PB was also assessed by optical, scanning, and transmission electron microscopies of liver done at days 60 and 120. Both Carcinosin 200 and Chelidonium 200 when administered alone show considerable ameliorative effect against p-DAB-induced hepatocarcinogenesis in mice; but the conjoint feeding of these two drugs appears to have had a slightly greater protection. These homeopathic remedies have the potential to be used as complementary and alternative medicine in liver cancer therapy, particularly as supporting palliative measures.”

My comments on Study 4:

 It is well known that liver tumors were induced in mice through chronic feeding of p-DAB and phenobarbital.This reported experiment has proved that molecular imprints contained in  carcinosin 200, as well as Chelidonium 200 can rectify the hepatocarcinogenesis induced by  p-dimethylaminoazobenzene (p-DAB)-induced hepatocarcinogenesis in mice. That means, some molecular imprints contained in carcinosin 200 and chelidonium 200 could bind to the ‘p-dimethylaminoazobenzene’ and ‘phenobarbital’ molecules, and remove the pathological inhibitions they produced in the enzyme systems involved in genetic expressions which caused the carcinogenic changes. This experiment also no way prove that potentized drugs “act” by  “interacting with DNA”- it only proved potentized drugs can prevent dna damages caused by the inhibitions of genetic expression enzyme systems produced by pathogenic molecules.

KHUDA-BUKHSH STUDY 5. Amelioration of carcinogen-induced toxicity in mice by administration of a potentized homeopathic drug, natrum sulphuricum 200- By Bhattacharjee NPathak SKhuda-Bukhsh AR.

“To examine if a potentized homeopathic drug, Natrum Sulphuricum 200 (Nat Sulph-200) has protective potentials against hepatocarcinogenesis,liver tumors were induced in mice through chronic feeding of P-dimethylaminoazobenzene (p-DAB; initiator of hepatocarcinogenesis) and phenobarbital (PB; promoter),  Mice were divided into five sub-groups: fed normal low protein diet (Gr. I, normal control); fed normal low protein plus alcohol-200 (vehicle of the homeopathic remedy) (Gr. II); fed diet mixed with 0.06% p-DAB plus 0.05% PB (Gr. III); fed diet and carcinogens like Gr.III, plus alcohol 200 (positive control for drug fed mice) (Gr. IV) and fed diet and carcinogens like Gr. III, plus Natrum Sulphuiricum-200 (Gr. V; drug fed). Mice were sacrificed at day 7, 15, 30, 60, 90 and day 120 for study of cytogenetical endpoints like chromosome aberrations (CA), micronuclei (MN), mitotic index (MI) and sperm head anomaly (SHA) and biochemical toxicity parameters like aspartate amino transferase (AST), alanine amino transferase (ALT), acid (AcP) and alkaline (AlkP) phosphatases, lipid peroxidation (LPO) and reduced glutathione (GSH) content. Less number of liver tumors were observed in Gr. V (drug fed) mice. Administration of Nat Sulph 200 reduced genomic damage, activities of AcP, AlkP, AST, ALT, LPO and increased GSH content. Therefore, independent replication of the study by others is encouraged.”

 My comment on Study 5

This study proved that molecular imprints contained in potentized  Natrum Sulph could remove the molecular inhibitions by P-dimethylaminoazobenzene and Phenobarbital. This experiment also no way prove that potentized drugs “act” by  “interacting with DNA”- it only proved potentized drugs can prevent dna damages caused by the inhibitions of genetic expression enzyme systems produced by pathogenic molecules.

KHUDA-BUKHSH STUDY 6. Can administration of potentized homeopathic remedy, Arsenicum album, alter antinuclear antibody (ANA) titer in people living in high-risk arsenic contaminated areas? I. A correlation with certain hematological parameters.- By Belon PBanerjee PChoudhury SCBanerjee ABiswas SJKarmakar SRPathak SGuha BChatterjee SBhattacharjee NDas JKKhuda-Bukhsh AR.

 “To examine whether elevated antinuclear antibody (ANA) titers reported in random human population of arsenic contaminated villages can be reverted to the normal range by administration of a potentized homeopathic drug, Arsenicum album, randomly selected volunteers in two arsenic contaminated villages and one arsenic-free village in West Bengal (India) were periodically tested for their ANA titer as well as various blood parameters in two types of experiments: ‘placebo-controlled double blind’ experiment for shorter duration and ‘uncontrolled verum fed experiment’ for longer duration. Positive modulation of ANA titer was observed along with changes in certain relevant hematological parameters, namely total count of red blood cells and white blood cells, packed cell volume, hemoglobin content, erythrocyte sedimentation rate and blood sugar level, mostly within 2 months of drug administration. Thus, Arsenicum album appears to have great potential for ameliorating arsenic induced elevated ANA titer and other hematological toxicities.”

MY comments on Study 6: 

This study scientifically proves molecular imprints contained in potentized forms of ars alb can act as artificial binding sites for arsenic ions and antidote the harmful biological effects produced by crude arsenic alb. This study do not or is not intended to provide any proof regarding the ‘biological mechanism’ of actions of potentized homeopathic drugs, but it proves ‘homeopathy works’.

KHUDA-BUKHSH STUDY 7. Evaluation of protective potentials of a potentized homeopathic drug, Chelidonium majus, during azo dye induced hepatocarcinogenesis in mice- ByBiswas SJKhuda-Bukhsh AR.

“Several cytogenetical and enzymatic protocols were used to test if two microdoses of Chelidonium majus, namely Chelidonium-30 (Ch-30) and Chelidonium-200 (Ch-200), used as homeopathic drugs, showed anti-tumor activity and also favorably modulated genotoxic damages produced by an azo dye in mice at several intervals of fixation. Different sets of healthy mice were fed: (i) hepatocarcinogen, p-dimethylaminoazobenzene (p-DAB, initiator) + phenobarbital (PB, promoter), (ii) only p-DAB, (iii) only PB, and (iv) neither p-DAB nor PB (normal control). Mice fed with p-DAB + PB were divided into different sets that were also fed either Ch-30 (v) or Ch-200 (vi) or diluted alcohol (vii), the “vehicle” of the microdoses of Chelidonium. All mice of group (i), a few of group (ii) and group (vii) and none of groups (iii) and (iv) developed tumors in liver at the longer intervals of fixation. The frequencies of chromosome aberrations (CA), micronucleated erythrocytes (MN), mitotic index (MI) and sperm head abnormality (SHA) were much higher in groups (i) and (vii) mice than in groups (ii), (iii) and (iv) mice at all fixation intervals. However, in mice of both groups (v) and (vi), the frequencies of CA, MN, SHA were strikingly less than those of groups (i) and (vii), and moderately less than those of groups (ii) and (iii). Both Ch-30 and Ch-200 also modulated favourably some toxicity marker enzymes like acid and alkaline phosphatases, peroxidases, glutamate oxaloacetate and glutamate pyruvate transaminases in liver, kidney and spleen tissues of the carcinogen fed mice. The microdoses of Chelidonium having no visible ill effects of their own, may be strong candidates for use in delaying/protecting liver cancer.”

My comments on Study 7:

 According to the authors themselves, this scientific study proves potentized Chelidonium has no specific  “anti-tumor activity”  against tumors  produced by hepatocarcinogens, ‘p-dimethylaminoazobenzene+ phenobarbital’ in mice”. This result indirectly proves that potentized drugs will have any effects only if indicated according to ‘similia similibus curentur’. It has to be particularly noted that Prof; Khuda Bukush is never concerned about this aspect of homeopathy while designing his research protocols. It also disproves his hypothesis of ‘genetic modulation’, since it shows ‘genetic modulation’ will happen only if the drug is homeopathically indicated by the presence of pathogenic molecules having configurational affinity towards the selected potentized drugs. It is evident from this study that potentized drugs act up on pathogenic molecules, not on genes as proposed by Prof Khuda Bukush. It is sad that  he failed to arrive at this conclusion.

KHUDA-BUKHSH STUDY 8. Comparative efficacy of pre-feeding, post-feeding and combined pre- and post-feeding of two microdoses of a potentized homeopathic drug, mercurius solubilis, in ameliorating genotoxic effects produced by mercuric chloride in mice- by Datta SBiswas SJKhuda-Bukhsh AR.

  “Mercury and its derivatives have become an alarming environmental problem, necessitating the search for effective antagonists, including homeopathic drugs, which are generally used in micro doses and are devoid of any palpable side-effects. On the basis of homeopathic similia principle, two potencies of Mercurius solubilis (Merc Sol-30 and Merc Sol-200) were tested by three administrative modes, i.e. pre-feeding, post-feeding and combined pre- and post-feeding, for their possible efficacy in ameliorating mercuric chloride-induced genotoxicity in mice. Healthy mice, Mus musculus, were intraperitoneally injected with 0.06% solution of mercuric chloride at the rate of 1 ml/100 g of body weight, and assessed for genotoxic effects through conventional endpoints. i.e. chromosome aberrations, micronuclei, mitotic index and sperm head abnormality, keeping suitable controls. Mercuric chloride-treated mice were divided into three sub-groups, which were orally administered with the drug prior to, after and both prior to and after injection of mercuric chloride, and their genotoxic effects were analysed at specific intervals of fixation. Mercuric chloride treatment generally produced more chromosome aberations, micronuclei and sperm head anomaly in mice, but the mitotic index appeared to be slightly reduced. While chromosome aberations, micronuclei and sperm head anomaly were generally reduced in the drug-fed series, the mitotic index showed an apparent increase. In most cases, the combined pre- and post-feeding mode appeared to show the maximum amelioration, followed by post-feeding and pre-feeding, in that order. The amelioration by Merc Sol-200 appeared to be slightly more pronounced. We conclude that potentized homeopathic drugs can serve as possible anti-genotoxic agents against specific environmental mutagens, including toxic heavy metals.”

 My comments on Study 8:

  It is known that mercury can produce ‘genotoxic’ effects by inhibitin certain enzymes involved in the processes of  dna replication and cell division and genetic expression, which could be observed by the conventional endpoints. i.e. chromosome aberrations, micronuclei, mitotic index and sperm head abnormality. In this experiment, it is proved that potentized forms of merc chrloride can prevent or reverse this harmful biological effects of mercury. That means, molecular imprints contained in potentized merc chroride can act as artificial binding sites to bind to mercury ions, and there be relieve the biological molecules from the inhibitions they produced. This experiment no way indicate potentized drugs “interact with genetic substance”, but it only proves that they “interact with” pathogenic molecules that have inhibited the biological molecules that underlie  “genotoxic” effects of mercury.

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